PCR-vector-sequencing

wetsel_r at wums.wustl.edu wetsel_r at wums.wustl.edu
Wed Dec 2 17:03:36 EST 1992


In a previous article, DMEIER at mis.mcw.edu wrote:
>Hello to all,
>  I am designing a set of primers which will give me a PCR product of
>approx. 600 bases.  These primers will have EcoR I and Hind III 
>restriction sites (one only for each primer of course).
>  Is there a plasmid of choice I should put the PCR product in for 
>sequencing?
>  All replies will be gratefully received.
> 
>Daniel A. Meier
>DMEIER at mis.mcw.edu
----------
Yes Dan....  Take your pick from the plethora of available cloning vectors. 
Personally, I'd take the old reliable p-bluescript.  1) it has both sites 
that you are using in its polyliker, 2) it has multiple primers for 
sequencing, 3) in HB101, JM109, and Sure (Stratagene) cells it is present 
in very high copy number, which translates into mgs/liter of t-broth or 
L-Broth, and 4) if sequencing is difficult for some reason, one can roll 
bluescript into a single stranded form for easier sequencing.

thats my $0.02 worth...

Best of luck,
David



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